AmpliTaq 360 DNA Polymerase amplifies a broad range of targets

Figure 1. Comparison of AmpliTaq 360 DNA Polymerase with other DNA Polymerases. Panel A shows data produced with AmpliTaq 360 DNA Polymerase while Panel B shows the same amplicons amplified using the Roche Taq DNA Polymerase. PCR reactions were performed using 1 ng of template DNA and 1.25 units of enzyme in each 50 uL reaction. Annealing was uniform across the selected targets. Annealing temperatures were uniform across the amplicon panel. Amplicons ranged from 300 to 1,400 basepairs in length with an average length of 553. Each reaction was performed in duplicate. The high GC target reactions included 2–10 uL of 360 GC Enhancer depending on the target used.

Figure 2. Comparison of AmpliTaq 360 DNA Polymerase with other DNA Polymerases. Panel A shows data produced with AmpliTaq 360 DNA Polymerase while Panel B shows the same amplicons amplified using the Stratagene Taq2000 DNA Polymerase. PCR reactions were performed using 1 ng of template DNA and 1.25 units of enzyme in each 50 uL reaction. Annealing was uniform across the selected targets. Annealing temperatures were uniform across the amplicon panel. Amplicons ranged from 300 to 1,400 basepairs in length with an average length of 553. Each reaction was performed in duplicate. The high GC target reactions included 2–10 uL of 360 GC Enhancer depending on the target used.

Figure 3. Comparison of AmpliTaq 360 DNA Polymerase with other DNA Polymerases. Panel A shows data produced with AmpliTaq 360 DNA Polymerase while Panel B shows the same amplicons amplified using the New England Biolabs Taq DNA Polymerase. PCR reactions were performed using 1 ng of template DNA and 1.25 units of enzyme in each 50 uL reaction. Annealing was uniform across the selected targets. Annealing temperatures were uniform across the amplicon panel. Amplicons ranged from 300 to 1,400 basepairs in length with an average length of 553. Each reaction was performed in duplicate. The high GC target reactions included 2–10 uL of 360 GC Enhancer depending on the target used.

Figure 4. Comparison of AmpliTaq 360 DNA Polymerase with other DNA Polymerases. Panel A shows data produced with AmpliTaq 360 DNA Polymerase while Panel B shows the same amplicons amplified using the Sigma RedTaq DNA Polymerase. PCR reactions were performed using 1 ng of template DNA and 1.25 units of enzyme in each 50 uL reaction. Annealing was uniform across the selected targets. Annealing temperatures were uniform across the amplicon panel. Amplicons ranged from 300 to 1,400 basepairs in length with an average length of 553. Each reaction was performed in duplicate. The high GC target reactions included 2–10 uL of 360 GC Enhancer depending on the target used.

Figure 5. Comparison of AmpliTaq 360 DNA Polymerase with other DNA Polymerases. Panel A shows data produced with AmpliTaq 360 DNA Polymerase while Panel B shows the same amplicons amplified using the Promega GoTaq Green. PCR reactions were performed using 1 ng of template DNA and 1.25 units of enzyme in each 50 uL reaction. Annealing was uniform across the selected targets. Annealing temperatures were uniform across the amplicon panel. Amplicons ranged from 300 to 1,400 basepairs in length with an average length of 553. Each reaction was performed in duplicate. The high GC target reactions included 2–10 uL of 360 GC Enhancer depending on the target used.

Specificity

Summary of 2 replicates for each of 40 amplicons indicating the average specificity for each amplicon and the standard deviation. AmpliTaq Gold® 360 PCR reactions were performed using 1 ng of template DNA per reaction using cycling conditions according to the manufacturer’s recommendations.

Annealing and extension times and temperatures were specific to each primer set. Amplicons ranged from 300 to 1400 base pairs (bp) in length with an average length of 553 bp.

Robust sequencing

Standard (A) and high GC % (B) amplicons were sequenced for both AmpliTaq® 360 DNA Polymerase and the original AmpliTaq®-generated PCR products.

10 ng of PCR products were used per sequencing reaction. Samples were cycle-sequenced using Big Dye® Terminator v3.1 and standard thermal cycling conditions. Big Dye XTerminator® was used to clean up the cycle sequencing reaction, and the samples were injected into the 3730xl sequencing instrument with POP-7 polymer.